【摘要】 目的 探究宣肺达郁汤含药血清对脂多糖(lipopolysaccharide,LPS)诱导的急性肺损伤A549细胞的影响及其作用机制。方法 将培养的人肺泡上皮细胞A549分为对照(Control)组、模型(LPS)组、宣肺达郁汤含药血清低剂量(XFDYL+LPS)组、中剂量(XFDYM+LPS)组、高剂量(XFDYH+LPS)组、地塞米松(DEX+LPS)组。其中Control组细胞不做干预，其余各组建立急性肺损伤炎症模型；Blank组、LPS组采用空白血清干预，其余各组采用相应含药血清干预。采用细胞计数试剂盒-8(cell counting kit-8,CCK8)法检测细胞增殖情况；采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测炎症因子[肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-8(interleukin-8,IL-8)、白细胞介素-6(interleukin-6,IL-6)]的含量；采用试剂盒检测氧化指标[谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)]的含量；采用实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测血管紧张素转换酶2(angiotensin converting enzyme 2,ACE2)、血管紧张素(1-7)[angiotensin(1-7),Ang(1-7)]、Mas mRNA的表达水平；采用蛋白印迹(western blotting)法检测ACE2、Ang(1-7)、Mas蛋白的表达水平。结果 与Control组相比，LPS组细胞的增殖能力显著降低，IL-6、IL-8、TNF-α、MDA表达水平显著上升(P<0.01),GSH水平显著降低(P<0.01),ACE2、Ang(1-7)、Mas的mRNA及蛋白表达量均显著降低(P<0.05)；与LPS组比较，宣肺达郁汤含药血清各组细胞增殖抑制率均显著升高，IL-6、IL-8、TNF-α、MDA表达水平均显著降低(P<0.05),ACE2、Ang(1-7)、Mas的mRNA及蛋白表达量均显著增加(P<0.05)。结论 宣肺达郁汤含药血清可能通过调控ACE2/Ang(1-7)/Mas通路抑制炎症因子释放、减轻氧化应激损伤，从而改善由LPS诱导的急性肺损伤A549细胞。更多还原

【Abstract】 Objective To investigate the effect of Xuanfei Dayu Decoction-containing serum on A549 cells with acute lung injury induced by lipopolysaccharide(LPS) and its mechanism. Methods The cultured human alveolar epithelial cells A549 were divided into Control group, model( LPS) group, low-dose(XFDYL+LPS), medium-dose(XFDYM+LPS), and high-dose(XFDYH+LPS) groups containing Xuanfei Dayu Decoction, and dexamethasone(DEX+LPS) group. The cells in the Blank group were not intervened, and the acute lung injury inflammatory model was established in the other groups; the Control group and LPS group were intervened with blank serum, and the other groups were intervened with corresponding drug-containing serum. The cell counting kit-8(CCK8) method was used to detect the cell proliferation. The enzyme linked immunosorbent assay(ELISA) method was used to detect the levels of inflammatory factors [tumor necrosis factor-α(TNF-α), interleukin-8(IL-8), and interleukin-6(IL-6)]. The kit was used to detect the levels of oxidation indexes [glutathione(GSH) and malondialdehyde(MDA)]. Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the expression levels of angiotensin converting enzyme 2(ACE2), angiotensin(1-7)[ Ang(1-7)], and Mas mRNA. Western blotting was used to detect the expression levels of ACE2, Ang(1-7), and Mas proteins. Results Compared with the Control group, the LPS group showed decreased cell proliferation and significantly increased expression of IL-6, IL-8, TNF-α, and MDA(P<0.01), while GSH was significantly decreased(P<0.01). The expression of ACE2, Ang(1-7), and Mas was also significantly decreased(P<0.05). Compared with the LPS group, the Xuanfei Dayu Decoction-containing serum groups showed increased inhibition rates and significantly decreased expression of IL-6, IL-8, TNF-α, and MDA(P<0.05). The expression of ACE2, Ang(1-7), and Mas was significantly increased(P<0.05). Conclusion Xuanfei Dayu Decoction-containing serum may alleviate acute lung injury in A549 cells induced by LPS by modulating the ACE2/Ang(1-7)/Mas pathway, inhibiting the release of inflammatory factors, and reducing oxidative stress damage.更多还原